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dc.date.accessioned2020-12-01T20:23:13Z
dc.date.available2020-12-01T20:23:13Z
dc.date.created2020-11-24T15:49:02Z
dc.date.issued2021
dc.identifier.citationAeschimann, Walter Kammer, Stephan Staats, Stefanie Schneider, Petra Schneider, Gisbert Rimbach, Gerald Cascella, Michele Stocker, Achim . Engineering of a functional γ-tocopherol transfer protein. Redox Biology. 2021, 38
dc.identifier.urihttp://hdl.handle.net/10852/81326
dc.description.abstractα-tocopherol transfer protein (TTP) was previously reported to self-aggregate into 24-meric spheres (α-TTPS) and to possess transcytotic potency across mono-layers of human umbilical vein endothelial cells (HUVECs). In this work, we describe the characterisation of a functional TTP variant with its vitamer selectivity shifted towards γ-tocopherol. The shift was obtained by introducing an alanine to leucine substitution into the substrate-binding pocket at position 156 through site directed mutagenesis. We report here the X-ray crystal structure of the γ-tocopherol specific particle (γ-TTPS) at 2.24 Å resolution. γ-TTPS features full functionality compared to its α-tocopherol specific parent including self-aggregation potency and transcytotic activity in trans-well experiments using primary HUVEC cells. The impact of the A156L mutation on TTP function is quantified in vitro by measuring the affinity towards γ-tocopherol through micro-differential scanning calorimetry and by determining its ligand-transfer activity. Finally, cell culture experiments using adherently grown HUVEC cells indicate that the protomers of γ-TTP, in contrast to α-TTP, do not counteract cytokine-mediated inflammation at a transcriptional level. Our results suggest that the A156L substitution in TTP is fully functional and has the potential to pave the way for further experiments towards the understanding of α-tocopherol homeostasis in humans.
dc.languageEN
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.titleEngineering of a functional γ-tocopherol transfer protein
dc.typeJournal article
dc.creator.authorAeschimann, Walter
dc.creator.authorKammer, Stephan
dc.creator.authorStaats, Stefanie
dc.creator.authorSchneider, Petra
dc.creator.authorSchneider, Gisbert
dc.creator.authorRimbach, Gerald
dc.creator.authorCascella, Michele
dc.creator.authorStocker, Achim
cristin.unitcode185,15,12,59
cristin.unitnameTeoretisk kjemi
cristin.ispublishedtrue
cristin.fulltextpostprint
cristin.qualitycode1
dc.identifier.cristin1851816
dc.identifier.bibliographiccitationinfo:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Redox Biology&rft.volume=38&rft.spage=&rft.date=2021
dc.identifier.jtitleRedox Biology
dc.identifier.volume38
dc.identifier.doihttps://doi.org/10.1016/j.redox.2020.101773
dc.identifier.urnURN:NBN:no-84407
dc.type.documentTidsskriftartikkel
dc.type.peerreviewedPeer reviewed
dc.source.issn2213-2317
dc.identifier.fulltextFulltext https://www.duo.uio.no/bitstream/handle/10852/81326/5/1-s2.0-S2213231720309782-main.pdf
dc.type.versionPublishedVersion
cristin.articleid101773
dc.relation.projectNFR/262695


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